Protein Expression

Ted Gauthier, Bollich, Patricia A.  |  8/20/2010 12:22:15 AM

Protein Expression Services

1)  Tobacco Expression System (Other plant systems are available.  Contact Ted Gauthier for more information.)

Expression of recombinant proteins in Nicotiana benthamiana by using an in planta based tobacoo mosaic virus system that allows very rapid and high levels of protein expression.  Recombinant Agrobacteria are delivered into tobacco plants by vacuum infiltration and recombinant tobacco mosaic virus is assembled expressing the protein of interest.

2)  E. coli Expression System

Expression of recombinant proteins in E. coli by using various commercially available vectors allows for very rapid and high levels of protein expression.  This simple, robust, low cost system is recommended for the production of antigen proteins, protein standards and functional proteins.

3)  Insect Cell Expression System

Invitrogen's Bac-to-Bac® expression system provides a rapid and efficient method of generating recombinant baculoviruses.  A choice of pFastBac donor plasmids allows easy construction of recombinant plasmids where expression of the gene of interest is controlled by a baculovirus-specific promoter.  An E. coli host strain, DH10Bac, which contains a baculovirus shuttle vector (bacmid) and a helper plasmid, allows generation of recombinant bacmid following transposition of the pFastBac expression construct.  After plasmid DNA purification, insect cells are transfected with a recombinant bacmid DNA, producing recombinant baculoviruses in the supernatant of infected Sf9 cells.  This method is designed to achieve virtually 100% recombination efficiency and recombinant protein expression is evaluated using recombinant baculovirus amplified without plaque purification in as early as passage 2.

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